Equipment at the Institute of Biochemistry and Signal Transduction

On this page we would like to inform you about the technical equipment of our institute.

We are dedicated to sharing our expertise and equipment. Please direct your requests to the secretariate of the institute (Kerstin Schluenz, k.schluenz@uke.de ).

Content

Analytical InstrumentsProtein and Sample PreparationCell Handling and Processing
Analytical HPLCFPLC SystemsEukaryotic Cell Culture
Mass SpectrometryLiquid Handling StationCell Electroporation
Light SpectroscopyMass Spectrometry Sample PreparationShaking Incubators
Multi-mode Plate ReaderLyophilization and Vacuum ConcentrationCell Disruption
Imaging Systems Superspeed and Ultracentrifugation
Real-time quantitative PCR  

Analytical Instruments

Analytical HPLC

Analytical HPLC

Prominence UFPLC
Shimadzu

Components / Specifications:
Autosampler
Binary gradient pump (up to 400 bar)
UV-Vis and Fluorescence Detector
Fractionator

Applications:
Protein binding studies via analytical Size Exclusion Chromatography (aSEC)
Nucleotide analysis and quantification (Ion-pairing RP-chromatography)
Reversed-Phase (RP-) Chromatography of proteins and peptides

Mass Spectrometry

Mass Spectrometry

amaZon speed LCMS
Bruker Daltonik

Specifications:
Type: 3D-Trap (Paul-Trap)
Detection range: m/z 50-3000 and m/z 200-6000
Resolution scan: 52,000 u/sec at 0.58 u FWHM
Electro Spray Ionization (Apollo II ESI-Source)
MS/MS-Mode: Fragmentation via CID
Coupled LC: Ultimate 3000 UHPLC (Thermo)

Selected Applications:
Detection of small proteins (up to 50 kDa)
Bottom-Up Proteomics via MS/MS-Analysis of peptides
Structure determination of organic compounds/metabolites

maXis II ETD LCMS (available from 08/2020)
Bruker Daltonik

Specifications:
Type: Ultra-High Resolution UHR-Q-TOF Analyzer
Quadrupole Isolation: up to 3000 m/z
Detection range: 50-40000 m/z
Full Sensitivity Resolution: 80,000 FSR
Electro Spray Ionization (ESI + NanoESI Source incl. NanoBooster dopant delivery)
MS/MS-Mode: Fragmentation via CID+ETD (parallel in the same run)
Coupled LC: Elute UHPLC (Bruker)

Selected Applications:
Mass determination of large intact proteins (up to 200 kDa)
Protein sequencing via Top-Down Mass Spectrometry of intact proteins
Mass Spectrometry of native proteins
Quantification or structure elucidation of organic compounds and metabolites (in DDA or DIA mode)

Light Spectroscopy

Fluorescence Spectrometer FP-8300
JASCO

Specifications:
Cuvette based device
Monochromators: 200 - 900 nm (Ex. and Em.)
Sensitivity: >8000:1 (RMS)
Accessories:
- Polarization filters for UV-Vis range
- Temperature-controlled single cell holder
- Magnetic stirrer
- A diverse set of quartz cuvettes

Selected Applications:
Fluorescence Intensity-based enzyme and substrate kinetics (FI)
Anisotropy-based binding studies via fluorescence polarization (FP)

Lambda 20

UV-Vis Spectrometer Lambda20
PerkinElmer

Specifications:
Cuvette based dual-beam device
Monochromators: 200 - 800 nm
Accessories:
- Single and multiple cell holders
- Temperature control (external)
- A diverse set of quartz cuvettes

Applications:
Time-resolved enzyme and substrate kinetics
Enzyme coupled assays
Absorption spectrum scanning

Multi-mode Plate Reader

Multi-mode Plate Reader

Spark
Tecan

Specifications:
Readout from 1well to 1536 well ANSI/SBS Plates
Temperature control: 18-45°C (long term stable)
Dual Optics: Filters and Monochromators
Measure Modes:
- Absorption UV-Vis: 200 – 1000 nm (Ex. and Em.)
- Fluorescence (FP + FI bottom and top; TRF; FRET)
- Luminescence – glow, flash, multicolor, and spectra
Accessories:
- Injectors – two channels min. 5 µL
- Automated lid lifter
- IR-Laser for ALPHA Technology
- Microscope: Bright Field Optics
- NanoQuant Plate / Humidity Chamber / Cell Chip adapter
- Absorbance cuvette port

Selected Applications:
Enzyme kinetic studies via optical readouts (e.g. Fluorescence Intensity; FI)
Anisotropy based binding studies via fluorescence polarization (FP)
24well cultivation screening platform (E. coli and Yeast)
High-throughput and low-volume DNA/RNA and protein quantification (2 µL)

Imaging Systems

BZ 9000 Fluorescence Microscope
Keyence

Specifications:
Resolution: 1360 x 1024 pixels
Bright Field (DIC)
Fluorescence Excitation Filters:
red (Cy5; 568 nm); green (GFP; 488 nm); blue (DAPI; 358 nm)
CO2- and heat-controlled chamber

Applications:
Imaging of slides and cell culture dishes (no Microtiter Plates) Scanning of small wells Multipoint Imaging Fluorescence Live Cell Imaging
Automated Cell Counting and Morphology Merge Measurements length / area / fluorescence (histograms)

Intas 2

ChemoStar Touch ECL & FL Imager
INTAS

Specifications:
6.0 MPixel CCD Camera
UV + Vis transmission
Enhanced Chemiluminescence (ECL)
Fluorescence detection (LED Cube RED 609nm)

Further ECL/FL Imager in-house:
LAS 3000 (Fujifilm)
Image Quant LAS 4000 (GE Healthcare)

Selected Applications:
Quantitative Western Blotting (WB) detection
SDS-PAGE and Agarose Gel documentation
Fluorescence detection in PAGE-gels

Real-time quantitative PCR

Applied Biosystems Quant Studio3
Thermo Scientific

Specifications:
96 Wells (reaction volume 10–100 μL)
Excitation/detection range 450–600 nm/500–640 nm
Multiplexing for up to 4 Targets
Three Independent Temperature Zones

Applications:
Quantification of gene expression levels (qPCR).
Differential Scanning Fluorometry (DSF) for indirect protein stability determination.

Analytical Instruments Accessible In-house:
IncuCyte Zoom Live Cell Analysis System (Sartorius)
CytoFLEX Flow Cytometer (Beckman Coulter)

Protein and Sample Preparation

FPLC Systems

BioRad Äkta

NGC Quest 10 Plus
BioRad

Specifications:
Piston Gradient Pumps (up to 252 bar, 25.2 MPa)
Flow Rates: 0.001–10 ml/min
Single Channel UV Detection at 280 nm
Separate Sample Pump
Fraction Collector BioFrac

ÄKTA Start
Ge Healthcare

Specifications:
Low-pressure peristaltic pump system
Single Channel UV Detection at 280 nm
Fraction Collector Frac30
Accessories: Electrophoresis System MiniPrep Cell (BioRad)

Further available FPLC-Devices: ÄKTA Prime plus (GE Healthcare)

Applications:
Protein purification via different chromatography strategies
- Affinity (IMAC, MBP-Tag, Strep-Tag, GST-Tag)
- Ion Exchange (Anion, Cation)
- Size Exclusion Chromatography (SEC), Desalting

Liquid Handling Station

epMotion

epMotion 5075t
Eppendorf

Specifications/Accessories:
15 deck positions
1- and 8-Channel Pipetting Tools (1 µL-1000 µL)
Transfer gripper
Thermomix Unit (up to 1400 rpm, 10-95°C)
2 temperature-controlled positions (2-100°C)
Magnetic separation module (96well format)

Selected Applications:
Semi-Automated DNA-Miniprep up to 96 samples in parallel
Automated small-scale recombinant protein purification
Automated MALDI mass spectrometry sample preparation
High Throughput Aliquotation (up to 192 reaction tubes in parallel)

Mass spectrometry sample preparation

Preparation hood

Clean Work Cabinet
Self-construction

Specifications / Accessories:
- Aspiration unit
- Centrifuge Minispin (Eppendorf)
- Airlock for sample transfer
- Magnetic racks

Applications:
Analytical workflows for MS-samples
Pulldown experiments with magnetic beads or centrifugation

Lyophilization and Vacuum Concentration

ALPHA 1-2 LD Freeze Dryer
Martin Christ

Specifications:
Temperature limit: -65°C
Vacuum limit <0.01 mbar (Pump RD-8; Vacuubrand)
Sample capacity: 8 Vacuum Adapters (for NS32/29 round flasks)
Vacuum glasses 600 mL for vials (0,5 mL – 50 mL)

ALPHA RVC Rotational Vacuum Concentrator
Martin Christ

Specifications:
Pressure regulation 1000 – 1 mbar
Rotational speed: up to 1300 rpm
Temperature regulation: 0 - 60°C
Accessory: Alpha 1-2 cooling trap + vacuum pump (Martin Christ)

Applications:
Lyophilization of proteins, peptides or chemicals
Quick vacuum concentration of liquid samples

Cell Handling and Processing

Mammalian Cell Culture

Equipment / Specifications:
Biological Safety Cabinets Type A2 in S2 Laboratories (Herasafe, Thermo)
CO2-Incubators (Heracell, Thermo)
Automated Fluorescence Cell Counter (Countess II FL, Thermo)

Application:
Cultivation of different mammalian cell lines and primary cells.
Lentiviral transfection of different eukaryotic cell lines.

Electroporation

NEON Transfection System
Invitrogen / Thermo Scientific

Specifications:
Specific biologically compatible pipette tip chamber
Transfection Capacity: 2 x 104 to 6 x 106 cells per reaction

Application:
High-efficiency transfection of extremely difficult cells like immune, primary, and stem cells.
Delivery of DNA, RNA or proteins into cells


Gene Pulser Xcell Electroporation Systems
BioRad

Specifications:
Cuvette based system:
main unit, CE module, PC module, and cuvette chamber
Capacitance: 10–500 V, 25–3275 µF in 25 µF increments
Output Voltage: 10–3000 V

Application:
Transformation of electro-competent microbial cells (bacterial and yeast) (PC module)
Transfection of eukaryotic cell lines with nucleic acids via electroporation (CE module)

Shaking Incubators

Shaking incubators

Multitron Standard/ Innova 44
Infors HT / New Brunswick

Specifications:
5 Shaking incubators
Capacity 6 x 5L Erlenmeyer flasks per incubator
Temperature control: 10-45°C
Shaking speed: 30-300 rpm
Time dependent programs

Application:
Multi-liter parallel cultivation of E. coli or yeast for
recombinant protein expression.

Cell Disruption

Cell lysis system

CF1 High-Pressure Cell Disruptor
Constant Systems

Specifications:
min. Sample Volume: 10 mL
Pressure Limit: 2700 bar (40 KPSI) (free selection)
Processivity: 100 mL/min

SFX250 Digital Sonifier
BransonSpecifications:
max. Power: 300 W (Amplitude 10-100%)
Sample Volume: 0,5 mL – 500 mL
Single Tip and 24-Tip high throughput Adapter

Applications:
Cell disruption of bacteria and yeast
Degassing of Buffer Solutions (Sonifier)

Superspeed and Ultracentrifugation

Sorvall LYNX 6000 Superspeed Centrifuge
Thermo Scientific

Specifications:
Max. Speed: 29,000 rpm
Max. RCF: 100,605 x g
Max. Volume: 6 x 1000 mL
Temperature control: -20°C to +40°C
Available Rotors: Carbon Fibrelite
- F9-6x1000 LEX; F12-6x500 LEX; F20-12x50 LEX

Further centrifuges in-house:
- Sorvall RC 5Cplus; Sorvall RC6plus (Thermo Scientific)
- Rotors: GSA; SS-34; SM-24

Applications:
Cell harvesting from up to 1 L cultures
Cell lysate clearing of up to 500 mL

Optima L-60 Ultracentrifuge
Beckman

Specifications:
Max. Speed: 60,000 rpm
Max. RCF: 485,000 x g
Available Rotors:
- 70Ti; 50Ti; 55.2TI; SW 40TI; VTi50

Further UC devices:
- L7-80 Ultracentrifuge (Beckman): Rotors see above
- Tabletop Ultracentrifuge OPTIMA-TL (Beckman)
Rotors: TLA100; TLA100.2

Applications:
Native protein purification
Cell compartment Isolation
Density gradient centrifugation